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OBJECTIVE@#To investigate the inhibitory effect of matrine on the proliferation of human non-small cell lung cancer (NSCLC) and explore the possible molecular mechanism.@*METHODS@#Cultured human NSCLC A549 cells were treated with 0.4, 0.8, 1.2, 1.6, and 2.0 g/L matrine for 24, 48 or 72 h. CCK-8 assay was used for measuring the changes in A549 cell viability. The morphological changes of the cells were observed under a fluorescence microscope, and flow cytometry was employed for analyzing the cell apoptosis. The effects of matrine and the PI3K specific inhibitor LY294002 (10 nmol/L) on AKT pathway and autophagy-related proteins in A549 cells were investigated using Western blotting.@*RESULTS@#Matrine significantly inhibited the proliferation of A549 cells in a time- and dose-dependent manner ( < 0.05). At the concentration of 1.6 g/L or higher, matrine caused obvious cell shrinkage and fragmentation and significantly increased floating cells; autophagy vacuoles could be observed in the cells after acridine orange staining. Within the concentrations range of 0.8-1.6 g/L, matrine time- and dosedependently increased the cell apoptosis. Treatment of the cells with 1.6 g/L matrine and 10 nmol/L LY294002 resulted in significantly lowered expressions of p-AKT and p-mTOR proteins and increased the expression of light chain 3B (LC 3B), an autophagy-related protein, as compared with those in the control cells ( < 0.05).@*CONCLUSIONS@#We demonstrate that matrine inhibits the proliferation and induces autophagy and apoptosis of A549 cells by deactivating AKT pathway, suggesting the potential of matrine as an anti-cancer agent for lung cancer.
Assuntos
Humanos , Alcaloides , Apoptose , Autofagia , Carcinoma Pulmonar de Células não Pequenas , Proliferação de Células , Neoplasias Pulmonares , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Quinolizinas , Transdução de Sinais , Serina-Treonina Quinases TORRESUMO
Objective To validate the feasibility of the simulationmethod and the reliability of thesimulationresult through comparison between simulation and measurement of the energy spectrum from medical diagnostic X-ray (RQR-Radiation qualities in radiation beans emerging from the X-ray source assembly).Methods A simplified model of the medical diagnostic X-ray RQR radiation quality was established using code of BEAMnrc.The energy spectrum of the same RQR radiation quality were measured through a plane high-purity germanium spectrometer,and compared with the simulationresult.Results The difference of spectral distribution between measurement and simulation was less than 3％,in spite of the convolution processing not happened to the pulse height distribution measured by the spectrometer.And the spectral distribution,fluence,energy fluence,means energy distribution of the radiation was obtained using the code of BEAMDP.Conclusions As indicated above,it is possible to use the simulation of the energy distribution as a foundation for the establishment of X-ray RQR radiation quality.
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Objective:To observe the conditions of isolation,purification and culture of mesenchymal stem cells(MSCs) in vitro derived from human umbilical cord,and to observe the repairing effect on experimental hepatic fibrosis in rats by transplantation with MSCs only or MSCs with IL-10.Methods:The MSCs derived from human umbilical cord were isolated,purified and cultivated in vitro.The model of hepatic fibrosis induced by CCl4 was established.After the model of hepatic fibrosis was succeeded,the rats were divided into three groups:model group,MSCs transplantation group,MSCs transplantation with IL-10 group.MSCs transplantation group was given 1×106 MSCs by the tail vein injection at the first 1 weeks,2 weeks and 3 weeks.MSCs transplantation with IL-10 group not only was given MSCs the same with MSCs transplantation group, but also was injected of IL-10 ( 4 μg/kg ) in abdomen, 4 times a week.After 4 weeks,the rats were executed,rats blood was collected to observe the changes of liver function and the index of liver fibro-sis.The liver HE staining was carried out to observe the changes of liver pathology.Expression of MMP-2 mRNA was detected by RT-PCR method,expression of TGF-βprotein was detected by Western blot method.Results:The rat liver function and the index of liver fibrosis improved obviously with a significant difference between model group , MSCs transplantation group and MSCs transplantation with IL-10 group after transplanting,MSCs with IL-10 was better than MSCs only.Liver tissue of rats with hematoxylin eosin staining suggested,liver fibrosis was obviously improved,and the MSCs transplantation with IL-10 group was best.MMP-2 mRNA expression was significantly higher between normal group and model group,MSCs transplantation group with a significant difference (P0.05).TGF-βprotein expression of other groups was higher than normal group with a significant (P<0.05),and MSCs transplantation with IL-10 group was the largest decrease than normal group.Conclusion:MSCs only or MSCs with IL-10 transplantation can improve the biochemical characteristics of rat peripheral blood and liver histological structure,and with IL-10 wan better than MSCs only.MSCs and IL-10 play a role in the treatment of liver fibrosis through lowering the expression of MMP-2 and the expression of TGF-βprotein.
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BACKGROUND:Recent studies verified that other tissues-derived stem cells can be homing to the liver, possibly participate in the regeneration of liver tissues, which provides new hope for stem cells in treatment of liver disease. OBJECTIVE:To observe isolation and culture of human umbilical cord-derived mesenchymal stem cells, to observe the repairing effect of umbilical cord-derived mesenchymal stem celltransplantation on hepatic fibrosis, and then to provide a reliable theoretical basis for further clinical application of umbilical cord-derived mesenchymal stem cells. METHODS:Human umbilical cord-derived mesenchymal stem cells were isolated and purified by natural adherent method, and then cultured and amplified in vitro. A rat model of hepatic fibrosis was prepared using subcutaneous multi-point injection of CCl4. A total of 22 rat models were randomly assigned to model injury group (n=11) and celltransplantation group (n=11). At 1, 2 and 3 weeks after model induction, the rats in the celltransplantation group were treated with 1×106 umbilical cord-derived mesenchymal stem cells via caudal vein. Four weeks later, the rats were sacrificed. Blood of rats was col ected from each group to detect liver functions. The liver was removed to receive hematoxylin-eosin staining, and pathological changes were observed. The number and distribution of Kupffer’s cells were observed using immunohistochemistry. The localization of umbilical cord-derived mesenchymal stem cells from treatment group was observed using immunohistochemistry.RESULTS AND CONCLUSION:After umbilical cord-derived mesenchymal stem cells were infused into rats with cirrhosis via caudal vein, liver function was significantly improved, which showed significant differences as compared with the control group (P<0.05). Hematoxylin-eosin staining revealed that hepatic fibrosis was apparently improved. Immunohistochemistry results demonstrated that the number of Kupffer’s cells was obviously reduced, and BrdU-labeled umbilical cord-derived mesenchymal stem cells were visible in rat liver of the treatment group using anti-BrdU antibody. These results suggested that umbilical cord-derived mesenchymal stem celltransplantation could improve biochemical characteristics of rat peripheral blood and histological structure of liver.
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<p><b>OBJECTIVE</b>To investigate the expression of beta-dystroglycan (beta-DG) and the roles of tissue inhibitor of metalloproteinases (TIMPs) on beta-DG in salivary adenoid cystic carcinoma (SACC).</p><p><b>METHODS</b>beta-DG in highly lung metastatic cell line ACC-M and lowly lung metastatic one ACC-2 was tested by immunocytochemistry with different concentrations (10, 15, 20, 25 micromol x L(-1)) of TIMPs, and that without the regulation of TIMPs was served as controls. beta-DG was detected in seven specimens of SACC and ten cases of normal salivary gland tissues which were considered as a comparison group by immunohistochemistry.</p><p><b>RESULTS</b>There was no positive beta-DG immune-staining at the ACC-2 and ACC-M cell lines without TIMPs in the cell culture. beta-DG expressed after the regulation of TIMPs. beta-DG expression was localized predominantly in basement membrane of the acinus, while the negative results were distributed in the carcinoma cells and around the cancer cell nests.</p><p><b>CONCLUSION</b>Beta-DG is widely expressed by transmembrane protein that plays important roles in connecting the extracellular matrix to the cytoskeleton, the fracture of this structure means that it is easy to invade and transfer, so restoration of beta-DG expression by TIMPs is considered to be critical for successful treatment of SACC.</p>